Search

Apoptotic cell death plays a pivotal role in the development and/or maintenance of several tissues including thymus. Deregulated thymic cell death is associated with autoimmune diseases including experimental autoimmune encephalomyelitis (EAE), a prototype murine model for analysis of human multiple sclerosis. Because Thy28 expression is modulated during thymocyte development, we tested whether Thy28 affects induction of EAE as effectively as antigen-induced thymocyte deletion using Thy28 transgenic (TG) mice. Thy28 TG mice showed partial resistance to anti-CD3 monoclonal antibody (mAb)-induced thymic cell death in vivo, as assessed by annexin V-expression and loss of mitochondrial membrane potential. The resistance to anti-CD3 mAb-induced cell death in Thy28 TG mice appeared to correlate with a decreased c-Jun N-terminal kinase phosphorylation and reduced down-regulation of Bcl-xL. Moreover, thymic hyperplasia was detected in Thy28 TG mice, although thymocyte development was unaltered. Development of peripheral lymphoid tissues including spleen and lymph nodes was also unaltered. Thy28 TG spleen T cells showed an increased production of IFN-γ, but not IL-17, in response to both anti-CD3 and anti-CD28 mAbs. Finally, Thy28 TG mice displayed accelerated induction of EAE as assessed by disease incidence, clinical score, and pathology following immunization with myelin oligodendrocyte glycoprotein compared with control WT mice. These findings suggest that modulation of Thy28 expression plays a crucial role in the determination of thymic cell fate, which may contribute to the development of EAE through proinflammatory cytokine production.

We studied the levels of immunoglobulins in colostrum, milk and sera from two common variable immunodeficiency (CVID) mothers (M1 and M2), and in sera from their newborn infants. During pregnancy they continued intravenous immunoglobulin therapy (IVIG). Antibody levels from maternal and cord blood collected at delivery and colostrum and milk, collected on the 3rd and 7th post-partum days, respectively, were analyzed. Although cord/maternal blood ratios of total immunoglobulins and subclasses, as well as specific antibodies differed between M1 and M2, both showed good placental transfer of anti-protein and anti-polysaccharide antibodies, despite lower cord/maternal blood ratios in M2. Anti-Streptococcus pneumoniae antibody avidity indexes were similar between paired maternal and cord serum. Both mothers' colostrum and milk samples showed only traces of IgA, and IgM and IgG levels in colostrum were within normal range in M1, whereas M2 presented elevated IgG and low IgM levels, when compared with healthy mothers. The study of colostrum and milk activity showed that they strongly inhibited enteropathogenic Escherichia coli adhesion in vitro. CVID patients must be informed about the relevance of regular IVIG administration during pregnancy, not only for their own health but also for their immune immature offspring. Breast-feeding should be encouraged as colostra from these CVID patients strongly inhibited E. coli adhesion to human epithelial cells thus providing immunological protection plus nutritional and psychological benefits for the infant.

In this report, we describe a patient who contracted fatal cryptococcosis after the induction of hemodialysis. A 76-year-old man was hospitalized to initiate hemodialysis. On admission, clinical findings showed no signs of any infections, and hemodialysis was inducted favorably. On the 6th hospital day he suddenly had a dyspnea and died from acute respiratory failure having a dyspnea for only 6 h. By microscopic examination at autopsy, we detected microemboli in the pulmonary capillary arteries caused by Cryptococcus and that the embolic source was a multiple-abscessed spleen. To the best of our knowledge, this is the first reported case of pulmonary capillary microembolism caused by cryptococcemia.

Background: The retinoblastoma (Rb) pathway, which governs cell cycle progression, is frequently genetically altered in cancer, causing deregulated expression of the E2F-1 transcription factor, which promotes DNA synthesis and cell cycle progression. Recent studies show that E2F-1 also participates in apoptosis induction in a p53 dependent or independent manner. Despite its crucial role and paradoxical effects on cell turnover, the function of E2F-1 in human cancer is unclear. Aims: To evaluate E2F-1 expression using immunohistochemistry in 43 surgically resected oesophageal squamous cell carcinoma (OSCC) specimens. Methods: This study analysed the association of E2F-1 with tumour cell proliferation and apoptosis and the upstream regulators modulating these processes, and its impact on patient outcome. Tumour cell proliferation and apoptosis were assessed as percentage of MIB-1 positive or apoptotic cells (MIB-1 labelling index (MI) and apoptotic index (AI)), respectively. Results: Entire specimens showed abnormal expression of one or more upstream regulators of pRb/E2F-1. Although E2F-1 positivity was not associated with the expression of upstream regulators, it showed a linear and positive correlation with MI but not AI. Patients with high MI, low AI, or high E2F-1 positivity had significantly shorter recurrence free survival. By multivariate analysis, high MI and low AI were independently associated with recurrence free survival, but E2F-1 was not. Conclusions: Increased cell proliferation and decreased apoptosis are associated with adverse prognosis in patients with OSCC. Although E2F-1 remains a controversial prognostic factor, its expression was closely associated with tumour cell proliferation and might influence clinical outcome, mainly via cell cycle progression.

Aims: The long-term effects of dietary diacylglycerol (DAG) on body fat and lipid metabolism were studied in patients undergoing hemodialysis (HD). Methods: Ten patients (seven males, three females) ranging in age from 40 to 64 years were enrolled. During the test period, 9.8g of DAG was ingested per day for 3 months. Results: Body mass index did not change throughout the study. The abdominal fat area measured by CT scan decreased significantly at 3 months, and increased significantly 3 months after completion of the DAG ingestion period. The serum composition of very low-density lipoprotein (VLDL) decreased significantly at 3 months and high-density lipoprotein (HDL) increased significantly at 3 months; these were determined using polyacrylamide gel electrophoresis. Serum lipoprotein (a) decreased significantly at 3 months. Conclusions: Our study showed that 3-month ingestion of DAG reduced the amount of abdominal fat and improved serum lipid profiles in free-living HD patients.

Aims: Several studies have reported that dysregulation of β catenin or k-ras mutation promotes cyclin D1 expression. This study investigated the relation between cyclin D1 expression and clinicopathological parameters in carcinoma of the ampulla of Vater (CAV), and also assessed the relation between increased cyclin D1 expression and β catenin/k-ras status in this series. Methods: Thirty CAVs were evaluated for cyclin D1 expression by immunohistochemistry in relation to patient clinicopathological features. Aberrant β catenin expression and k-ras mutation were also investigated by immunostaining and direct sequencing, and related to cyclin D1 expression. Results: Increased cyclin D1 expression was seen in 17 of 30 CAVs and was significantly correlated with tumour cell proliferation and disease free survival time (p = 0.018, p = 0.018, respectively). Nuclear accumulation of β catenin was found in nine of 30 cases, including four cases with missense mutations in exon 3 of CTNNB-1, and was significantly correlated with increased cyclin D1 expression (p = 0.003). k-ras gene mutation was detected in 12 of 30 cases, and was also significantly correlated with increased cyclin D1 expression (p = 0.026). Overall, 14 of 17 CAVs with increased cyclin D1 expression showed nuclear accumulation of β catenin and/or k-ras mutation. Conclusions: Increased cyclin D1 expression appears to be associated with tumour proliferation and poorer clinical outcome in CAV. It is also associated with both aberrant β catenin expression and k-ras mutation. These results are consistent with the in vitro data that cyclin D1 can be transactivated by activated β catenin–T cell factor/LEF and k-ras pathways.

9-(4′-Aminophenyl)-9H-pyrido [3,4-b] indole (aminophenylnorharman, APNH) is a novel mutagenic heterocyclic amine, produced by the reaction of norharman with aniline in the presence of S9 mix. In the present study, the maternal and developmental toxicity of APNH were investigated in ICR mice administered oral doses of 0, 0.625, 1.25, 2.5 or 5 mg/kg/day on gestational days (GD) 6 through 15 or 0, 5, 10, or 20 mg/kg on GD 12. Maternal and foetal parameters were evaluated on day 18 of gestation. Foetuses of dams treated on GD 6–15 were examined for external and skeletal malformations and variations, and foetuses of dams treated on GD 12 were inspected for cleft palate. Maternal death occurred when APNH was administered at 5 mg/kg/day on GD 6 –15. No significant decrease in body weight gain during the administration period was observed at doses of 2.5 mg/kg/day or less when applied on GD 6– 15. Adverse changes in general condition of dams were observed in the groups treated at doses of 2.5 mg/kg/day and above on GD 6 –15, whereas no adverse effects on dams were noted even when APNH was applied at a fairly high dose on GD 12. Intracytoplasmic vacuolation in hepatocytes, necrosis of proximal tubular epithelial cells and desquamation of necrotic epithelial cells in the tubular lumen were observed in dams treated with APNH at 2.5 or 5 mg/kg/day on GD 6 –15. Increased pre-implantation loss was observed at 5 mg/kg/day and post-implantation loss was observed at 2.5 mg/kg/day and above when applied on GD 6–15, or at 20 mg/kg when applied on GD 12. Foetal body weight was decreased by APNH in a dose-dependent manner. The frequency of external malformations (cleft palate) was significantly increased in the group treated with APNH at 2.5 mg/kg/day on GD 6–15 compared to the controls. However, there were no foetuses with cleft palate even when APNH was given at 20 mg/kg on GD 12. No significant increases in skeletally malformed foetuses were found in any APNH-treated group. The frequency of lumbar ribs was increased dose dependently. This study demonstrated the develop-mental toxicity of a mutagenic compound, APNH, in mice at maternally toxic doses, and that cleft palate observed in term foetuses resulted from the adverse effect of APNH on the maternal environment during organogenesis. More detailed studies are warranted to assess the possible risks to pregnant women from exposure to APNH. Human & Experimental Toxicology (2002) 21, 147 – 151.

Background: Fanconi`s anemia (FA) is a rare recessive chromosomal instability disorder, characterized by progressive bone marrow failure and congenital defects. Patients with FA present with recurrent infections, particularly those of the respiratory tract. Objective: The aim of the present study was to evaluate whether patients with FA have altered antibody-mediated immune responses. Methods: A group of 12 patients with FA, 5–32 years old (6 males) was studied. Serum levels of IgG, IgM, IgA and IgG subclasses, isohemagglutinin titers and specific IgG antibodies to poliovirus and measles were determined using standard methods. Immediate skin tests to common inhalant allergens were performed, and total and specific serum IgE was quantitated using a fluoroenzymatic assay (Uni-CAP, Pharmacia). Antipneumococcal antibodies were measured by ELISA before and 4–8 weeks after immunization with pneumococcal vaccine (Pneumo 23, Pasteur Mérieux Connaught). Responses to serotypes 1, 3, 5, 6B, 9V and 14, which are the most prevalent in our country, were studied. Results: Ten patients had elevated IgE levels in sera, and 7 of them had detectable specific IgE and positive immediate skin tests. An inadequate response to pneumococcal vaccination was found in 2 of the 12 patients. Isohemagglutinin titers and levels of IgG, IgM, IgA and IgG subclasses and antipoliovirus and antimeasles antibodies were within the normal limits for age in all patients. Two patients had undetectable IgG4 levels (below 5 mg/dl). Conclusions: The results indicate that a proportion of patients with FA (2/12) in our study had inadequate responses to pneumococcal vaccination. No other significant abnormalities of the immune system were found in these patients.

In 40 children with Haemophilus influenzae b (Hib) meningitis, we determined serum levels (mg/dl) of IgG subclasses using the radial immunodiffusion method; 67.8 per cent of these children were less than 24 months old. In 14 children of the sample we measured serum IgG and IgG2 anti-ribosyl-ribitolphosphate (anti-PRP) (by enzyme-linked immunosorbent assay, ELISA) in the acute and convalescent phases of the disease. Lower IgG2 levels than those of the control group were obtained in all age ranges: 3-12 months, 1-2 years (p < 0.01), and 2-5 years (p < 0.001). IgG4 was also present in lower levels in patients of all age ranges (p < 0.05, p < 0.001, and p < 0.01 respectively). Serum levels of IgG anti-PRP and IgG2 anti-PRP measured were very low in the acute phase of the disease in all age ranges and there was no notable increase in levels during the convalescent phase of the disease. This result indicates that children less than 24 months old do not produce sufficient levels of IgG and IgG2 anti-PRP even after Hib meningitis.

1. The pharmacological properties of (R)-(+)-3-phenyl-5-{2-(1-pyrrolidinylmethyl)butyryl}isoxazole hydrochloride (MS-322) were investigated and compared with those of other muscle relaxants. 2. MS-322 (3–12 mg/kg, IV) reduced spinal reflex potentials in acutely spinalized rats dose dependently. Its inhibitory effects on polysynaptic and dorsal root reflexes were more marked than those on the monosynaptic reflex. 3. MS-322 (1.5–12.0 mg/kg, IV) depressed the afferent discharges of de-efferented muscle spindles in anesthetized rats dose dependently. 4. The inhibitory effect of MS-322 on this afferent activity was considered to be due to its membrane stabilizing action. 5. In addition to the central effects of MS-322, its effect on muscle spindles may contribute to its muscle relaxant activity.

We analysed data from 27 patients with herpes simplex virus (HSV) infections of the central nervous system (CNS) found in a 1990–1992 survey in Kyushu and Okinawa, Japan. Patients ranged in age from one year to 70 years, with peaks seen in the 20s and 50s. Temporal lobe-limbic encephalitis was the most common HSV infection (13 patients), followed by meningitis (5), diffuse encephalitis (4), disseminated encephalomyelitis (ADEM) (3) and brain stem encephalitis (2). Another three patients with non-herpetic, non-paraneoplastic acute limbic encephalitis were presented. Our study indicates that HSV infection can cause ADEM, although temporal lobe-limbic encephalitis or meningitis are more common. The early diagnosis of HSV-related ADEM is important because of the efficacy of the timely administration of corticosteroids.

We examined metabolic and functional changes when forebrain ischemia was induced in stroke-prone spontaneously hypertensive rats by bilateral carotid artery occlusion. In addition, the protective effect of clentiazem was evaluated in this model. Rats were anesthetized with urethane. Cerebral blood flow was measured with a laser Doppler flowmeter. Cerebral high-energy phosphates and intracellular pH were measured by phosphorus magnetic resonance spectroscopy. Electroencephalographic activity was evaluated as the summation of its amplitude. These parameters were monitored during a 30-minute period of ischemia and recirculation. Clentiazem was given orally as pretreatment (10 mg/kg twice a day for 3.5 days). Bilateral carotid occlusion caused a decrease in cerebral blood flow to approximately 5% of the preischemic level and the disappearance of electroencephalographic activity. Occlusion also caused a decrease in ATP and phosphocreatine (to 48.7 +/- 4.3% and 23.7 +/- 2.2% of preischemic levels, respectively) as well as intracellular pH (from 7.3 +/- 0.1 to 6.0 +/- 0.1). During recirculation the reversal of these changes was variable: high-energy phosphates were partially restored, but electroencephalographic activity and intracellular pH showed little improvement. Hypoperfusion (55.7 +/- 11.5% of the preischemic flow) developed after reactive hyperemia. Pretreatment with clentiazem lessened the decrease in cerebral blood flow (control, 4.8 +/- 1.4%; clentiazem, 14.1 +/- 4.1% of the preischemic level; P < .05) and prevented the disappearance of electroencephalographic activity in some rats during ischemia. Clentiazem also prevented postischemic hypoperfusion and accelerated the restoration of high-energy phosphates, intracellular pH, and electroencephalographic activity during recirculation. Carotid artery occlusion induced stable forebrain ischemia in stroke-prone spontaneously hypertensive rats. Clentiazem improved the metabolic and functional disturbances that occurred in this ischemic model, and its beneficial effect appeared to be due mainly to the relative preservation of cerebral blood flow during carotid occlusion.

Endothelium-dependent relaxations that are resistant to inhibitors of nitric oxide synthase probably are mediated by endothelium-dependent hyperpolarization of the vascular smooth muscle. Experiments were performed to examine the distribution of this type of relaxation along the arterial tree of the rat by measuring changes in isometric force. Acetylcholine induced concentration- and endothelium-dependent relaxations in aortas and in pulmonary, common iliac, femoral, mesenteric, and renal arteries contracted with phenylephrine. In the presence of NG-nitro-L-arginine, the cumulative administration of acetylcholine induced relaxations only in the femoral, mesenteric, and renal arteries. The calcium ionophore A23187 relaxed mesenteric arteries contracted with phenylephrine in a concentration- and endothelium-dependent manner. The concentration-relaxation curve to A23187 was shifted to the right in the presence of NG-nitro-L-arginine. The maximal relaxations induced by lemakalim, a K+ channel opener, were smaller in those arteries that did not exhibit NG-nitro-L-arginine-resistant relaxations. These results suggest that NG-nitro-L-arginine-resistant relaxations are more frequently observed in smaller arteries. The arteries that exhibit NG-nitro-L-arginine-resistant relaxations may be more sensitive to an endothelium-derived substance that causes hyperpolarization of vascular smooth muscle cells.

Nebivolol is a new β1-antagonist that acutely reduces arterial blood pressure without depressing cardiac function. The present study was designed to determine the effect of nebivolol on coronary arteries. Rings of canine left anterior descending coronary (LAD) artery with or without endothelium were suspended in organ chambers and the isometric tension was recorded. In some experiments, the transmembrane potential of the smooth muscle cells was recorded by electrophysiological method. During contractions to prostaglandin F2α, nebivolol induced concentration-dependent relaxations of the coronary arteries. The enantiomer. l-nebivold, also induced comparable relaxations; however, d-nebivolol induced smaller relaxations. The relaxations induced by nebivolol and its enantiomer were significantly larger in tissues with than in those without endothelium. The differences between tissues with and without endothelium were abolished by nitro-l-arginine (3 x 10-5M) or methylene blue (10-5M). The nebivolol-induced relaxations were not affeced by indomethacin (10-5M), phentolamine (5 x 10-6M), propranolol (5 x 10-6M), or methysergide (3 x 10-6M. Nebivolol at a subthreshold concentration for inducing relaxation (3 x 10-) did not significantly affect endothelium-dependent relaxations to acetylcholine but potentiated ADP-induced endothelium-dependent relaxations. The potentiation is stereoselective for l-nebivolol. Nebivolol induced a small hyperpolarization of the coronary smooth muscle with endothelium (1 mV). These observations demonstrate that (a) nebivolol induces relaxations of canine coronary arteries that are mainly mediated by endothelium-dervied relaxing factor(s): these effects are not related to the α- and β-adrengergic- or serotonegergic 5-HT1- and 5-HT2 blocking properties of the compound; (b) nebivolol potentiates ADP-induced endothelium-dependent relaxations: and (c) l-nebivolol is more potent than d-nebivolol in inducing and potentiating endothelium-dependent relaxations.

Background. In Japan, living donor renal transplantation has gained momentum due to an increased number of patients with end-stage renal disease. Living donation not only provides better outcomes, but also the recipients usually need less medications, thereby increasing the quality of life and reducing the potential side effects of immunosuppression. Materials and Methods. For the past 25 years, our center had performed 140 open donor nephrectomy (OPNx) renal transplantations. Since July 2003, we changed our procurement operation to living hand-assisted laparoscopic donor nephrectomy (HALNx) in 49 cases. Our operative technique consisted of two 12-mm ports placed in the midaxillary line at the superior and inferior levels of the umbilicus. Next, a 5-cm incision was made in the midline periumbilicus and the hand port system fitted through a midline abdominal incision. Results. In 49 cases, HALNx was completed successfully; no patient required conversion to laparotomy. The estimated blood loss was 33.0 ± 43.4 g and no patient required blood transfusion. In comparison, in OPNx the blood loss was 426.5 ± 247.6 g (P

Background and purpose: Sorbin and SH3-domain-containing-1 (SORBS1) is an important adaptor protein in insulin-signalling pathway, and its genetic polymorphism may regulate the activity of insulin resistance. We investigated the association between the SORBS1 T228A polymorphism and ischaemic stroke. Methods: Genotyping was achieved by a rapid-cycle PCR and melting curve analysis using fluorescent probes in 1049 incident cases of ischaemic stroke and 1049 age- and sex-matched control subjects recruited from the Hisayama study. Results: The allele distributions of the SORBS1 T228A polymorphism were similar amongst cases and controls. The multivariate-adjusted odds ratio (OR) of the AA genotype for ischaemic stroke was 2.897 (95% CI, 0.907–8.018) compared with the TT genotype. In terms of stroke subtype, there was a trend toward a difference in the AA genotypes for lacunar infarction, compared with the TT genotype (OR = 8.740, P = 0.0510), and combined TT and TA genotypes (OR = 8.768, P = 0.0505). The other polymorphisms genotyped were not associated with any subtypes of ischaemic stroke. T228A polymorphism of SORBS1 was not associated with the prevalence of diabetes. Conclusions: The AA genotype of SORBS1 T228A polymorphism may play a role in lacunar infarction in the Japanese population.

Male ICR mice were intraperitoneally injected with TEM, MMC, ENU, PCZ, or PMS and mated to untreated females of the same strain on days 64–80 after the treatment. Copulations during this period involve sperm that were spermatogonial stem cells at the time of the treatment. The fetuses were examined on day 18 of pregnancy for external and skeletal abrnomalities. The 5 mutagnes tested all caused significant increases in the incidence of abnormal fetuses over the control level. The genotoxically effective dose, in mmole/kg, for producing fetal abnormalities with a frequency of 2% was estimated to be 0.007 for TEM and MMC, 0.6 for ENU, 1.8 for PCZ, and 3.0 for PMS. These values correlate well with the mutagenic potency estimated from the data reported for inducing specific-locus mutations in spermatogonial stem cells. Irrespective of the kind of mutagen used, external abnormalities represented by cleft palate and dwarfism occurred more frequently than skeletal abnormalities represented by rib malformations. It is concluded from these data that F 1 fetal abnormalities can serve as sensitive indicators for quantitatively assessing the genotoxicity of a chemical agent in spermatogonial stem cells.

In our department, there were 313 thoracic surgeries for primary lung cancer from January 1994 to December 2003. We clinically reviewed for the operative and hospital death (n=18, 5.8%). The patients were 16 males and 2 females (70.6 +/- 5.6 years old). The surgical procedures were 4 pneumonectomies, 13 lobectomies (3 bronchoplasties) and 1 partial resection. The mean interval until postoperative death was 122.5 +/- 156.1 days. There were 5 direct operative deaths within 30 days (1.6%). There were 4 cancer deaths, 2 hemoptyses, 2 operative bleeding, 2 thromboses, 2 cerebral hemorrhages, 1 pyothorax, 1 pneumonia, 1 respiratory failure, 1 multiple organ failure after chemotherapy and 2 unexplained deaths. The patients with pneumonectomy or aged significantly had high mortality. For postoperative complications such as hemoptysis or bleeding, perioperative management that takes these issues into consideration is needed. Furthermore, we must carefully review the preoperative evaluation and combined treatment, because there were many cancer deaths among cases showing early recurrence and metastasis.

Since radiation and chemotherapy have little impact on survival and no prospect for cure, surgery offers the best potentially option in patients with liver tumors. However, a lot of patients with liver tumors is not resectable due to stage combined with health problems, or poor liver function reserve. In this study, our preliminary clinical reports in patients with unresectable liver tumors treatment with RFA have demonstrated radiologic evidence of tumor necrosis and complications. Multielectrode, radiofrequency probes were supplied by RITA Medical Systems (Mountain View, California). This study involved a total of 28 liver tumors in 19 patients (including 17 patients with hepatomas and 2 patients with metastases). The mean age was 68.9 years old. The size of tumor was more than 3 cm in 15 patients. The approaches to the tumor were laparoscopic in 1 patients and open surgical in 18 patients. Hepatic vascular occlusion was combined with RFA in 5 cases. The reasons for unresectable were defined as total bilirubin, ICGR 15, cardiopulmonary function and multiplicity. The efficacy of this RFA therapy was evaluated by preoperative and follow-up CT scans or tumor makers. There tumor necrosis was shown in 15 patients (78.9%). The survival rates at the time up to 12 months were 84.2% in 16 patients. The present study has demonstrated that the RFA is able to provide a safe and effective means in controlling liver tumors.

Three neurodegenerative diseases causing primary autonomic failure are pure autonomic failure (PAF), Parkinson's disease (PD), and multiple system atrophy (MSA). Differential diagnoses among these diseases are often difficult especially in early disease stage. For example, it may be difficult to determine whether a patient with parkinsonism and autonomic failure has PD or MSA. Recently, a decrease in myocardial uptake of meta-iodobenzylguanidine (MIBG), an analogue of norepinephrine, has been reported in PD but not in MSA using [123I]MIBG myocardial scintigraphy.1 This new imaging approach is thought to be of significance in the diagnosis and characterisation of akinetic rigid syndromes, especially PD. After that, we reported severe loss of cardiac sympathetic nerves in one patient with PD but not in one patient with MSA, which accounts for a difference in myocardial uptake of MIBG between PD and MSA.2 However, our observation was based on the study in only a single patient of each disease. In this study, we immunohistochemically examined the heart tissues from four patients with PD, three patients with MSA, and one patient with PAF, and showed the involvement of postganglionic cardiac sympathetic nerves in PD and PAF …

Although the pathogenesis of oral lichen planus (OLP) is not clear, a small proportion of cases with OLP are reported to transform to cancer. We examined the epithelial cell proliferation status of OLP to relate the labelling index to microscopic features surveyed routinely in pathology. Mucosal biopsies obtained from 44 cases diagnosed with OLP with an intact oral epithelium and 10 normal control specimens from Japanese subjects were immunohistochemically stained with MIB and p53 antibodies. The Ki67 labelling index (LI) was significantly higher in OLP compared with normal controls. A particularly large number of OLP lesions (64%) were p53 positive. No association was, however, found with p53 expression and the Ki67 LI. Atrophic and flat epithelia had a quantitatively higher LI, which did not significantly differ from acanthotic biopsies. Increased cell proliferation in OLP is likely to be a secondary phenomenon due to the damage inflicted on keratinocytes by infiltrating mononuclear cells in the submucosa.

The susceptibility of 129/SvJ mice to infection with Strongyloides venezuelensis was compared with that of C57BL/6 mice. After a primary infection, daily egg output in faeces (EPG) from 129/SvJ mice was lower and terminated earlier than that from C57BL/6 mice. Adult worm recovery from the small intestine of 129/SvJ mice on day 7 was also lower than that of C57BL/6 mice. When the numbers of larvae recovered from the lungs were examined on days 2, 3 and 4 after a primary infection, they were comparable between the two strains. On the other hand, when an equal number of larvae recovered from the lungs of each strain on day 3 were implanted orally into homologous strain mice, the magnitude of EPG and the number of adult worms in the small intestine on day 5 after implantation were significantly lower in 129/SvJ than in C57BL/6 mice. The number of mucosal mast cells in the jejunum was not significantly different between 129/SvJ and C57BL/6 naive mice. Total chondroitin sulphate concentration in the gut washings obtained from naive mice was significantly higher in 129/SvJ (11.34 +/- 9.48) than in C57BL/6 mice (1.09 +/- 0.77, P0.05). These results indicate that the natural resistance of 129SvJ mice to S. venezuelensis infection is expressed at the intestine, probably due to higher concentration of chondroitin sulphate, which prevents establishment of S. venezuelensis.

The aim of this study was to elucidate any association between oral lichen planus (OLP) and serum antioxidant micronutrients by a population-based case-control study. A total of 9536 subjects were examined, and 62 (58 reticular, 4 atrophic and erosive) diagnosed with OLP at referral facilities were compared with four controls per case (n=248) selected among disease-free subjects matched for age and sex. Serum levels of micronutrients (retinol, alpha-tocopherol, zeaxanthin/lutein and cryptoxanthin, lycopene, alpha-carotene and beta-carotene) were estimated by the high performance liquid chromatography (HPLC) method. Among cases, mean serum retinol level (2.820+/-0.849 micromol/l) was significantly higher compared with that of controls (2.562+/-0.735 micromol/l) (P0.05). No significant differences were noted in carotenoid levels examined in this study, except for a lower level of lycopene found in atrophic/erosive OLP cases. The results of this study suggest that low serum retinol or carotenoid levels are not risk factors for occurrence of lichen planus, and any specific benefits of antioxidant micronutrients cannot be claimed for this inflammatory disorder.

The present study was designed to investigate the contribution of membrane hyperpolarization to endothelium-dependent relaxations induced by serotonin in the porcine coronary artery.Rings with and without endothelium of porcine coronary arteries were suspended in conventional organ chambers for the measurement of isometric force. The cell membrane potential of the vascular smooth muscle cells was measured using glass microelectrodes, in the presence of indometacin, ketanserin, and/or N omega-nitro-L-arginine.Serotonin induced a transient endothelium-, and concentration-dependent relaxation in rings contracted with prostaglandin F2 alpha in the presence of N omega-nitro-L-arginine (maximal relaxation: 19%). The N omega-nitro-L-arginine resistant relaxation was abolished by high K+ and tetrabutylammonium chloride. Serotonin also caused an endothelium-, concentration-dependent membrane hyperpolarizations with a maximal amplitude of -8.8 mV. The nitro-L-arginine resistant relaxations and hyperpolarizations were abolished by methiothepin, but not by glibenclamide. The time course of the endothelium-dependent relaxations and hyperpolarizations was similar.These results suggest a contribution of cell membrane hyperpolarization to the endothelium-dependent relaxations induced by serotonin in the porcine coronary artery.

To elucidate whether a relationship exists between the site of trauma and severity of acute hyperextension spinal cord injury without bone damage, we examined the clinical features of 25 male and 10 female patients aged 13 to 88 years. None of the patients had vertebral damage such as fracture and dislocation. The site of impact was classified as the buccal, forehead, or mandibular region. The neurological findings were assessed according to Frankel's classification at admission and at follow up after 3 months or more to assess outcome. Eleven patients suffered trauma in the buccal region, one patient in Frankel's grade B, three in grade C, and seven in grade D at admission. All 11 of these patients showed an improvement of one grade or more to an outcome of C in one patient, D in one, and E in nine. Trauma occurred at the forehead region in 18 patients, four in grade B, 10 in grade C, and four in grade D. Improvement was seen at follow up by one grade or more to C in one patient, D in 10, and E in seven. Trauma occurred at the mandibular region in six patients, four in grade B and two in grade C. Four of these patients showed improvement of one grade or more to grade B in one, grade C in four, and grade E in one. Overall, seven patients had poor outcomes, five of whom suffered trauma to the mandibular region, indicating that impact to the mandibular region tends to have an unfavorable clinical outcome. Our findings indicate that the site of trauma greatly influences the severity of hyperextension spinal cord injury.

We propose the designation 'sialolipoma' to establish and characterize a new category of benign lipomatous tumour occurring in salivary glands. Until now, these tumours have not been regarded as a distinct entity in the salivary glands.We evaluated the clinicopathological and immunohistochemical features of seven sialolipomas among 2051 surgically resected primary salivary gland tumours deposited in our files. The seven patients with sialolipoma were five men and two women, aged 20-75 years (mean: 54.4 years). Five tumours had arisen in the parotid gland, one in the soft palate, and one in the hard palate. The tumours ranged from 10 to 60 mm (mean: 38 mm) in maximum diameter. Histologically, the tumours were characterized by a well circumscribed mass composed of glandular tissue and mature adipose elements. The adipose elements in the tumours arising in the parotid gland were more abundant than those arising in the minor salivary gland. The glandular components consisted of ductal, acinar, basal and myoepithelial cells, and closely resembled the cellular and structural compositions of normal salivary gland tissues. These findings were confirmed by immunohistochemical and ultrastructural studies. These components had no atypia, except for the presence of some minor variations, e.g. ductal ectasia with fibrosis and focal oncocytic metaplasia. In all cases, cell proliferative activity, as assessed by Ki67 (MIB1) immunostaining, was low. From these findings, it is likely that our cases were lipomas with secondary entrapment of the salivary gland elements. No recurrence was seen in all cases after superficial parotidectomy, or after surgical excision in the patients with palatal tumours.We regard sialolipoma as a distinct variant of salivary gland lipoma that can occur in both the major and minor salivary glands. Superficial parotidectomy, or surgical resection in the case of palatal tumours, is an appropriate treatment for this benign tumour.

We report the results of a phase III trial comparing tacrolimus (FK506) with cyclosporine for GVHD prophylaxis after allogeneic BMT. From February 1995 to July 1996, 136 patients were enrolled and followed up to September 1997. During the first 100 days post-transplant the incidence of grade II-IV acute GVHD (the primary end-point) was lower in the tacrolimus group (17.5%) compared with the cyclosporine group (48.0%, P < 0.0001). A significant difference was observed between the tacrolimus and cyclosporine groups when subset analyses were performed based on recipients from HLA-matched siblings (13.3% vs 41.3%, P = 0.015) or donors other than HLA-matched siblings (21.4% vs 53.8%, P= 0.0029). The incidence of chronic GVHD (47.3% and 47.8%) and Kaplan-Meier estimate of overall survival (62.9% and 65.2%) were similar between the tacrolimus and cyclosporine groups, respectively. The overall leukemia relapse rate was not significantly different between the tacrolimus and cyclosporine groups (19.6% and 11.4%, respectively). However, the relapse rate among recipients from HLA-matched siblings was significantly higher in the tacrolimus group (30.9%) compared with the cyclosporine group (3.6%, P = 0.013). These results suggest the merit of tacrolimus for the prophylaxis of acute GVHD, but a lack of merit for a graft-versus-leukemia effect among recipients from HLA-matched sibling donors.

A large difference in immunosuppressive potency between methylprednisolone and prednisolone has been suggested in vitro. However, the selection of the best glucocorticoid for renal transplantation has been seldom considered so far. Thus, the present study was undertaken to compare therapeutic efficacy between prednisolone and methylprednisolone in renal transplantation. We studied 42 renal transplant recipients who were operated on between 1990 and 1994. The patients were divided into two treatment groups: a methylprednisolone/ cyclosporine group (n = 19) and a prednisolone/cyclosporine group (n = 23). Clinical outcome and drug side effects were compared retrospectively between the treatment groups 24-84 months after transplantation. The overall graft survival time in patients treated with methylprednisolone/cyclosporine was superior to that in patients treated with prednisolone/cyclosporine (p0.05). Among the recipients from cadaver donors, 5/16 (31.3%) treated with prednisolone required nephrectomy, whereas none of the 10 patients treated with methylprednisolone received nephrectomy (p0.01). An examination of the recipients from living related donors revealed that serum creatinine levels 24-36 months after operation were significantly lower in the methylprednisolone group (p0.05). Cyclosporine trough levels and glucocorticoid side effects were similar between the treatment groups. The results raised the possibility that methylprednisolone is superior to prednisolone when combined with cyclosporine for maintenance immunosuppressive therapy in renal transplantation.

Stereotactic radiosurgery (SRS) for brain metastasis results in a high local control rate. But cystic metastatic tumor should have been a contraindication for SRS. Because it is often found that a cyst is too large to be irradiated, the tumor does not exist in the center of the irradiation field. Between 1995 and 1998, 8 consecutive patients underwent linear accelerator-based SRS for cystic brain metastases identified by computed tomography or magnetic resonance image scan. Stereotactic cyst aspiration is carried out after placement of the BRW frame under local anesthesia. All of the patients except one were confirmed to have sufficient reduction of the cysts. 5-7 days after stereotactic cyst aspiration, SRS was performed. The dose range was 25-30 Gy. In follow-up MRI, local recurrences and enlargement of cysts were not noted. Six patients with neurological symptoms recuperated satisfactorily. Median survival was 30 weeks from the date of radiosurgery. All of the patients died and the causes of death were related with the primary lesion. We conclude that our technique, a combination of stereotactic cyst aspiration and SRS is an effective measure which leads to palliation of neurologic symptoms and is a low risk treatment for patients with cystic brain metastasis.

Intraductal papilloma is an extremely rare benign salivary gland tumor that occurs most commonly in the minor salivary glands. To our knowledge, a malignant counterpart of intraductal papilloma has not been described previously. We report one case each of benign and malignant intraductal papillary tumors. The benign tumor occurred in the sublingual gland and was a typical example of intraductal papilloma, with the exception that we found no previously published reports of this type of tumor in this location. The other patient had a left parotid gland tumor that was architecturally similar to the intraductal papilloma, with the addition of cytologic atypia, intraductal extension, microinvasion, and lymph node metastases. This tumor was diagnosed as intraductal papillary adenocarcinoma with an invasive component. Both patients were alive and well without evidence of recurrence 2 years and 6 months (case 1) and 6 years (case 2) after surgery. Immunohistochemical examination revealed that the tumor cells resembled duct luminal cells in both cases. The 2 tumors had different immunoreactivities for carcinoembryonic antigen, p53, and Ki-67. The malignant counterpart of intraductal papilloma should be considered in the differential diagnosis of salivary gland tumors with a predominantly papillary structure, even though this tumor is extremely rare.

Measurement of 99mTc-MAG3 plasma clearance (CLmag) based on one-compartment model (MPC method) was applied to renal transplantation and evaluated for the factors which might affect the calculated results, especially concerning renal depth. Correlation coefficient of CLmag between MPC method using real renal depth and Russell or Bubeck single sampling method was good (r = 0.852 or 0.876, respectively). Regression equation between MPC method and Russell method was y = 1.044x - 3.0 and was more closer to y = x than that between MPC method and Bubeck method. CLmag of MPC method calculated by estimated renal depth from the abdominal thickness was also similar to that by real renal depth. Even if the fixed renal depth, 4 cm, was applied, the coefficient and regression equation between MPC method and Russell method were r = 0.884 and y = 1.004x - 10.2. In conclusion, MPC method is applicable to the evaluation of renal transplants. Though measuring renal depth is best, calculation with fixed renal depth of 4 cm might be practically acceptable.

Association between lymphocyte-sensitivity to immunosuppressants in transplant recipients in vitro and clinical outcomes has been demonstrated. In general, renal transplant recipients are treated with a combination of immunosuppressants such as either glucocorticoid/cyclosporin A (CsA) or glucocorticoid/tacrolimus (FK506) but the pharmacological complementarity of these drugs is still controversial. We examined relationships between the lymphocyte-sensitivities to these immunosuppressants.We examined lymphocyte-sensitivities to prednisolone (PSL), CsA, and FK506 in vitro in a total of 190 chronic renal failure (CRF) patients and 140 healthy subjects. The lymphocyte-sensitivity was evaluated from the IC50 value against mitogen-stimulated lymphocyte-blastogenesis in vitro.Statistically significant correlations of the IC50 values in CRF patients between the following pairs of drugs were observed: PSL and CsA (P0.0001; n=129, r=0.419), PSL and FK506 (P0.001; n=54, r=0. 441), and CsA and FK506 (P0.0001; n=45, r=0.608). Similar correlations were also observed in lymphocytes from healthy subjects. The population of CRF patients who exhibited high IC50 values (low sensitivities) to PSL and FK506 was significantly larger than that of healthy subjects (P0.05).Patients who showed low lymphocyte-sensitivity to either of the drugs also may exhibit low sensitivity to the others, and thus they may have a high risk of unsatisfactory outcome under combination therapy after renal transplantation. To overcome this risk, the selection of immunosuppressants is recommended to be restricted according to individual lymphocyte-sensitivities to these drugs in vitro, or alternatively, by addition of other drugs with different mechanisms for immunosuppression.

Variable clinical courses have been recognized in patients with idiopathic pulmonary fibrosis (IPF), but the disease is known to have a fundamentally unfavorable outcome. In addition, we have yet to fully understand whether the regional or population differences are negligible or not. Therefore, we analyzed the clinical outcomes in Japanese IPF patients in a hospital-based historical cohort study.A questionnaire was used to collect records for retrospective analysis of the IPF patients. We analyzed data collected from 51 hospitals and clinics in Japan on 234 Japanese IPF patients (males 170, females 64) with IPF who had been followed up after diagnosis.1. Five and ten years after the detection of exertional dyspnea, the overall survival rates were 41.0% and 20.1%, respectively. A higher age at the time of detection (over 59) was related to a decrease in survival rates. 2. The incidence of complications was no higher in the corticosteroid-treated cases than in the untreated cases. The effects of corticosteroid treatment on survival was not confirmed in this type of study.The survival rates of histologically proven Japanese patients with IPF were similar to the rates previously reported in different populations. Age at the detection of exertional dyspnea was critical in terms of the survival rate.

Coronary artery bypass grafting (CABG) on a beating heart has been successfully performed for high risk patients, and is known to be less invasive than conventional CABG using cardiopulmonary bypass (CPB). We expanded the indication of beating-heart CABG in patients requiring emergency coronary revascularization.A retrospective chart review was performed for patients who had undergone emergency CABG on a beating heart (EM-BH group), elective CABG on a beating heart (Elective-BH group) and emergency CABG under CPB (EM-CPB group), between January 1, 1997 and June 30, 1998.Four cases (1 male and 3 females with a mean age of 67.8 +/- 5.4) in the EM-BH group, 67 cases (48 males and 19 females with mean age of 67.3 +/- 7.8) in the Elective-BH group, and 41 cases (29 males and 12 females with mean age of 63.3 +/- 10.4) in the EM-CPB group were analyzed. The number of the grafts was 1.75 +/- 0.50 in EM-BH group, 1.37 +/- 0.55 in the Elective-BH group, and 2.95 +/- 1.07 in the EM-CPB group. The intubation period, ICU stay, and the postoperative hospital stay were significantly shorter in the EM-BH group (6.0 hours intubation, 1.5 days ICU stay, and 11.5 days postoperative hospital stay) and Elective-BH group (6.8 +/- 11.0 hours intubation, 1.6 +/- 1.5 days ICU stay, and 12.7 +/- 5.2 days postoperative hospital stay) than in the EM-CPB group (20.1 +/- 22.5 hours intubation, 3.6 +/- 2.4 days ICU stay, and 21.8 +/- 14.9 days postoperative hospital stay).The postoperative recovery period for EM-BH patients was almost the same as that for elective cases of beating-heart CABG, and was significantly shorter than that of conventional emergency CABG under CPB. Selected patients with coronary ischemia can be safely treated by beating-heart surgery.

A 16-year-old male experienced a sudden attack of back pain while walking through the corridor of school which required emergent hospitalization. Except for the back pain, no neurological symptoms were noted. Magnetic resonance (MR) imaging indicated an angiopathy-like flow void in the epidural region at Th 3-5 which seemed to explain the patient's back pain. Thoracic laminectomy at Th 3-5 and resection of the affected site were performed. Pathologically, the resected lesion only had a dilated normal vein and no findings indicating vascular deformity. The patient's outcome was good and no relapse of pain has occurred for about 2 years since the operation. Although some authors have reported vascular deformity with spinal epidural hemorrhage or varices with lumbar hernia of the intervertebral disc, there is no report concerning spinal epidural varices with pain only. The present case seemed to be a rare event and is reported here.

l-cis Diltiazem, an optical isomer of diltiazem, protects against myocardial dysfunction in vitro, whereas its Ca2+ channel blocking activity is about 100 times less potent than that of diltiazem. However, there is no evidence that l-cis diltiazem actually protects against ischemia/reperfusion injury in vivo. To assess this, we employed an anesthetized rabbit model, where the left circumflex artery was occluded for 15 min and reperfused for 360 min. Treatment with diltiazem before and during ischemia (bolus 200 microg/kg and 15 microg/kg per minute for 25 min, i.v.; 575 microg/kg total) showed slightly depressed hemodynamic parameters, while l-cis diltiazem (1150 microg/kg) had no effect. Treatment with l-cis diltiazem produced a high recovery of the thickening fraction and limited the infarct size in a dose-dependent manner. Furthermore, the treatment with l-cis diltiazem (1150 microg/kg) or diltiazem (575 microg/kg) 5 min before reperfusion also limited the infarct size, but not after reperfusion. These results suggest that l-cis diltiazem affects some events in the onset of reperfusion, independently of Ca2+-channel-blocking action. Our observations are the first to show that l-cis diltiazem demonstrated its cardioprotective action in the ischemic rabbit heart in vivo.

Although current chemotherapeutic regimens cure as many as 70% of children with acute lymphoblastic leukemia (ALL), infants continue to show a poor outcome. In this paper, we describe the outcome in 37 ALL infants treated between 1989 and 1995 in Japan. Patients had characteristic findings of infant ALL, including hyperleukocytosis100 x 10(9)/l (15/37, 41%), blast cells with a CD10-negative phenotype (30/37, 81%), and 11q23/MLL involvement (21/37, 57%). Seven were treated according to Aggressive Treatment Research Group protocol, 15 according to the Ministry of Health and Welfare protocol, and 15 according to protocols of other institutions. The 3-year overall event-free survival (EFS) was 33%. The EFS was 13% for infants aged26 weeks at diagnosis and 43% for infants aged26 weeks. Infants who had blast cells with CD10 negative phenotype with 11q23/MLL involvement were also associated with poor prognosis. However, infants with CD10 positive blasts without 11q23/MLL involvement had a better outcome (EFS 75%). These results suggest that intensive chemotherapy is effective for patients with good prognostic factors, but for infants with poor prognostic factors a more aggressive approach such as stem cell transplantation might be necessary.

Cystic partially differentiated nephroblastoma (CPDN) is an uncommon renal multicystic tumour, usually affecting early infants. To our knowledge, this report describes the first case of CPDN occurring in an adult.A 45-year-old man was found incidentally to have a left renal cystic tumour, measuring 20 mm in diameter, at the lower pole far from the pelvis. The tumour was composed of multilocular cystic spaces of variable size and intervening septa without solid nodular areas. The cysts were lined by a single layer of flattened, hobnail, or columnar epithelium. The septa were made of mesenchymal cells, which were admixed with small numbers of loosely aggregated blastemal cells, occasional tubular structures in various stages of development, and a few glomeruloid structures. The tumour cells had no anaplasia, and mitoses were rare. Immunohistochemical and lectin histochemical studies revealed that the cyst lining epithelium and the tubular structures in the septa expressed predominantly the markers for distal tubules and collecting ducts. Ultrastructurally, the cyst lining cells closely resembled collecting duct cells while some tubular structures showed an immature nephrogenic morphology. The patient was alive and well without evidence of recurrence 11 months after surgery.CPDN does occur in adults, as experienced in Wilms' tumour, though its incidence is extremely low. This study suggests that CPDN may show maturation intermediate between cystic nephroma and Wilms' tumour, even in adult cases.

We investigated temporal differences in the protective action of three types of Ca2+ channel blockers in myocardial ischemia, focusing particularly on the blocking ability under depolarizing conditions. The effects of diltiazem, verapamil, and nifedipine on extracellular potassium concentration ([K+]e), acidosis, and level of metabolic markers were examined during 30-min global ischemia and postischemic left ventricular (LV) function in isolated guinea pig hearts. Diltiazem and verapamil, but not nifedipine, inhibited the late phase (15-30 min) of [K+]e elevation, whereas all three blockers delayed the onset of the early phase (0-8 min) of [K+]e elevation. Diltiazem and verapamil inhibited ischemic contracture and improved postischemic LV function to a greater extent. These differences appeared to be linked to preservation of ATP and creatine phosphate and delay of cessation of anaerobic glycolytic activity. Maneuvers to preserve energy sources during ischemia (decrease in external Ca2+ concentration or pacing at a lower frequency) attenuated the late phase of [K+]e elevation. Inhibition of LV pressure was potentiated 12- and 8.2-fold by diltiazem and verapamil, respectively, at 8.9 mM K+ as compared with 2.9 mM K+, whereas that by nifedipine was unchanged. These results indicate that the differential cardioprotection of Ca2+ channel blockers in the late period of ischemia correlates with preservation of high-energy phosphates as a result of different Ca2+ channel blocking abilities under high [K+]e conditions.

Malignant transformation of Warthin's tumour (WT) is a rare event. We present two cases of mucoepidermoid carcinoma (MEC) arising in WT in the parotid gland.Two cases of MEC arising in WT, which were found in 185 cases of WT of the parotid gland, were investigated by light and electron microscopy, and immunohistochemistry. Both cases had largely similar macroscopic and histological features with some differences. Histologically, the tumours consisted mainly of WT with multilayered hyperplastic arrangements of oncocytic cells and focal squamous and goblet cell metaplasia. In the same tumour mass, however, the features of MEC were observed with invasion to adjacent adipose tissue. A transitional zone between WT and MEC was evident. Both patients were alive and well without evidence of recurrence 30 and 34 months after surgery, respectively. Electron microscopy revealed that cyst lining epithelial cells in WTs had abundant mitochondria whereas some of the MEC cells contained numerous tonofilaments and mucinous granules. Immunohistochemically, oncocytic cells of WTs were strongly positive for mitochondria and SalylTn was extensively stained in MECs. The labelling index for Ki67 was obviously higher in the carcinoma cells than the epithelial cells of WT.Our cases confirmed the possibility of malignant transformation of the epithelial component in WT to MEC.

In this report, we discuss the findings of a 1994 retrospective study concerning the rate of inhibitor formation in Japanese hemophiliacs. The study was the second of its kind, following on the first in 1991 (Kamiya et al. 1995 Int. J. Hematology). The records of 77 medical institutions were examined. Inhibitors were found in 6.50% (140 of 2154) of the patients with hemophilia A (HA), and 5.21% (22 of 422) of those with hemophilia B (HB). The median age for antibody formation was 10.7 years in patients with HA, and 4.5 years in those with HB. The median period (exposure period) from initial plasma factor concentrates exposure to inhibitor formation was 46 days and 20 days, respectively, in the HA and HB patients. Among the HA inhibitor patients, those with a large deletion or a nonsense mutation were aged 17.4 years or less (0.58, 1.7, 3.5, 5.5, 7.0 and 17.4), whereas those with intron 22 inversion were aged 55.7 years or less (1.3, 1.3, 1.8, 33.0, 36.1, 37.7, 43.9, 47.9 and 55.7).

Basal cell adenocarcinoma (BCAC) of the salivary gland is a rare tumor and recently described entity. Eleven cases of BCAC are presented here and compared with basal cell adenoma (BCA) through assessment of cell proliferative activity, apoptosis, and expression of p53, bcl-2, and epidermal growth factor receptor (EGFR) because these two tumors show close similarity in some cytologic and architectural characteristics.Formalin fixed, paraffin embedded sections of 11 cases of BCAC and 9 cases of BCA, selected from the authors' files of 1851 primary tumors of the major salivary gland, were examined using immunostaining for Ki-67 (MIB-1), p53, bcl-2, and EGFR. In addition, apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling.The incidence of BCAC was 0.6% among patients with major salivary gland tumors in the current series. Nine cases of BCAC arose in the parotid gland and two were of submandibular gland origin. Approximately 50% of the patients had recurrences, but no patient developed metastases or died of disease. Vascular involvement (75%), perineural invasion (36%), and necrosis (45%) were common features. Cell proliferative activity, including mitotic count, Ki-67 labeling index (LI), and apoptotic index were significantly higher in BCAC than BCA. More than four mitotic figures per ten high-power fields or a Ki-67 LI5% appeared to be limited to cases of BCAC. Considering those cases expressing p53 or EGFR in10% of tumor cells as positive, 6 of the 11 BCAC cases were positive for p53 and 3 were positive for EGFR. In contrast, all BCA cases were negative for p53 and EGFR. Although all cases of BCA were strongly positive for bcl-2 (50% of tumor cells), 3 of 11 cases of BCAC were completely negative.BCAC is a rare salivary gland tumor with a relatively high recurrence rate. Examination of cell proliferation, apoptosis, and expression of p53, bcl-2, and EGFR were found to be useful in distinguishing malignant basal cell tumors from their benign counterparts arising in the salivary gland.

To examine the contribution of beta-adrenoceptor (beta AR) downregulation to desensitization of beta ARs by chronic administration of a beta AR agonist, we compared the adenylyl cyclase (AC) activities in two kinds of cardiac ventricular membranes with decreased available beta ARs: one was derived from rats infused with a selective beta 1 AR agonist, T-0509 [(-)-(R)-1-(3,4-dihydroxyphenyl)- 2-[(3,4-dimethoxyphenethyl)-amino]ethanol hydrochloride], in vivo (40 micrograms/kg/hr, s.c. for 6 days); and the other was obtained from treatment of control membranes with an irreversible beta AR antagonist, bromoacetyl alprenolol methane (BAAM). T-0509 infusion decreased the densities of beta 1 ARs and beta 2 ARs by 26% and 32%, respectively, and reduced the maximal isoproterenol-stimulated AC activity by 53%. The amount of Gs alpha and Gi alpha proteins in the membranes was not significantly changed by T-0509 infusion. To make preparations that mimic the T-0509-induced downregulation, we treated the control membranes with 100 nM BAAM in vitro. The BAAM treatment decreased the Bmax value of [125I]iodocyanopindolol for beta 1 ARs and beta 2 ARs by 29% and 36%, respectively, whereas it reduced the maximal effect of isoproterenol on AC activity only by 37%. These results suggest that downregulation of beta ARs cannot fully account for the desensitization by chronic treatment of T-0509 and that other mechanism(s) can play a significant role in the loss of responsiveness.

A novel derivative of diltiazem (1,5-benzothiazepine Ca++ antagonist), DTZ323, 3-(acetyloxy)-5-[2-[[2-(3,4-dimethoxyphenyl)ethyl]-methylamino]eth yl]-2,3-dihydro-2-(4-methoxyphenyl)-1,5-benzothiazepine-4(5H)-one, was characterized by radioligand binding experiments with rabbit skeletal T-tubule membranes in terms of the affinity and the selectivity to the binding sites for the three classical calcium antagonists, such as dihydropyridines, phenylalkylamines and benzothiazepines. DTZ323, like diltiazem and clentiazem, exhibited complete and concentration-dependent inhibition of d-cis-[3H]diltiazem binding to the membrane with a slope factor close to unity. Ki values indicated that DTZ323 (Ki = 6.6 +/- 0.6 nM, mean +/- S.E., n = 4) was 48 times and 9 times more potent than diltiazem and clentiazem, respectively. DTZ323 partially inhibited the specific binding of a dihydropyridine ligand, (+)-[3H]PN200-110, at 37 degrees C. The equilibrium saturation study showed that DTZ323 reduces the affinity for the (+)-[3H]PN200-110 binding in a concentration-dependent manner with a slight decrease in the density of the binding sites. DTZ323 also inhibited the specific binding of a phenylalkylamine ligand, (-)-[3H]D888, completely as did diltiazem. DTZ323 (1 microM) had no effect on the dissociation rate of d-cis-[3H]diltiazem at 2 degrees C, whereas 30 microM verapamil increased the dissociation rate, which suggested that DTZ323 inhibits the specific binding of d-cis-[3H]diltiazem in a manner similar to other competitive ligands for the benzothiazepine binding site. These results indicate that DTZ323 is a selective ligand for the 1,5-benzothiazepine binding site with the highest affinity among the diltiazem derivatives.

1. We tested the hypothesis that activation of large conductance calcium-activated potassium channels is involved in dilator responses of the basilar artery to acetylcholine in vivo. Using a cranial window in anaesthetized rats, we examined responses of the basilar artery to acetylcholine. 2. Topical application of acetylcholine (10(-6) and 10(-5) M) increased diameter of the basilar artery from 238 +/- 7 microns to 268 +/- 7 and 288 +/- 7 microns, respectively (P0.05 vs. baseline diameter). Iberiotoxin (10(-8) M), an inhibitor of large conductance calcium-activated potassium channels, did not affect baseline diameter of the basilar artery. In the presence of 10(-8) M iberiotoxin, 10(-6) and 10(-5) M acetylcholine increased diameter of the basilar artery from 239 +/- 7 microns to 246 +/- 7 and 261 +/- 7 microns, respectively. Thus, iberiotoxin attenuated acetylcholine-induced dilatation of the basilar artery (P0.05). 3. Sodium nitroprusside (10(-7) and 10(-6) M) increased diameter of the basilar artery from 242 +/- 9 microns to 310 +/- 12 and 374 +/- 13 microns, respectively (P0.05 vs. baseline diameter). In the presence of iberiotoxin (10(-8) M), sodium nitroprusside (10(-7) and 10(-6) M) increased diameter of the basilar artery from 243 +/- 6 microns to 259 +/- 9 and 311 +/- 12 microns, respectively. Thus, iberiotoxin attenuated dilator responses of the basilar artery to sodium nitroprusside (P0.05). 4. Iberiotoxin partly inhibited dilator responses of the basilar artery to forskolin, a direct activator of adenylate cyclase, but did not affect vasodilatation produced by levcromakalim, a potassium channel opener. 5. These results suggest that dilator responses of the basilar artery to acetylcholine and sodium nitroprusside are mediated, in part, by activation of large conductance calcium-activated potassium channels. Because both acetylcholine and sodium nitroprusside have been shown to activate guanylate cyclase via nitric oxide, activation of large conductance calcium-activated potassium channels may be one of the major mechanisms by which cyclic GMP causes dilatation of the basilar artery in vivo.

Cord blood stem cell transplantation (CBSCT) was performed on a patient with acute promyelocytic leukemia. The patient was a boy 3 years and 8 months old, who had shown complete remission following treatment with intensive chemotherapy. However, after the final course of consolidation chemotherapy, chromosome analysis of his bone marrow aspirates revealed 46XY, t(15,17)(q22;q21), and a PML-RAR alpha fusion gene was detected by the reverse transcriptase-polymerase chain reaction test. All-trans retinoic acid diminished the chromosomal abnormality, but the PML-RAR alpha fusion gene remained. The patient was then treated with CBSCT from an HLA-matched sibling donor. The number of nucleated cells in the cord blood was 2.2 x 10(7)/kg of body weight, and that of granulocyte-macrophage colony-forming units 0.6 x 10(4)/kg. Methotrexate was given, on days 3 and 6, as prophylaxis against graft-versus-host disease (GVHD). The neutrophil count rose to above 500/microliters on day 22. The platelet count exceeded 50,000/microliters on day 48. Platelet transfusions were given 12 times after CBSCT, the last one on day 36. Grade I acute GVHD was treated with steroids. The patient was well and discharged on day 103, without symptoms or laboratory data suggestive of relapse. Following this experience we instituted a project of the Kanagawa Cord Blood Bank, which is scheduled for expansion nationwide.